丹皮酚对人胶质母细胞瘤细胞中血管内皮生长因子表达的影响

Paeonol Reduces Vascular Endothelial Growth Factor Expression in Human Glioblastoma Cells In Vitro

目的研究丹皮酚(paeonol,Pae)对人胶质母细胞瘤细胞U251中血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的影响。方法以实时定量PCR法检测VEGF转录水平,以Western blot检测VEGF表达水平。结果 1Pae可呈剂量依赖性下调U251细胞中VEGF的转录水平,当Pae作用浓度分别为47、94、188、376、752和1 504μmol/L时,对U251细胞VEGF mRNA的抑制率分别为29.6%、43.94%、73.78%、79.49%、83.11%和88.06%;2Pae对U251细胞VEGF转录的抑制作用随着时间的延长而逐渐增强,在188μmol/L浓度下,当Pae作用时间分别为4、12和24h时,对U251细胞VEGF mRNA的抑制率分别为67.13%、71.33%和82.04%;3Pae可显著降低U251细胞VEGF蛋白的表达,以47、188、376、1 504μmol/L浓度的Pae分别作用U251细胞24h后,VEGF蛋白的表达水平量明显降低,并呈剂量依赖性减弱,以上浓度Pae对U251细胞VEGF蛋白表达的抑制率分别为33.09%、73.93%、81.23%和87.85%。结论 Pae可显著降低人胶质母细胞瘤细胞中VEGF的转录及表达,有可能通过下调VEGF表达而干预人胶质母细胞瘤细胞的血管新生及化疗敏感性。

Objective To investigate the regulative effects of paeonol on the expression of vascular endothelial growth factor（VEGF）in human glioblastoma cells（U251）.Methods Real-time quantitative PCR was performed to detect the level of VEGF mRNA.The expressive level of VEGF protein was determined by Western blot.Results ①Paeonol down-regulated the transcription level of VEGF in U251 cells in a dose-dependent manner.The inhibition rate of VEGF mRNA level was 29.6%,43.94 %,73.78 %,79.49 %,83.11 % and 88.06 %,respectively,when U251 cells were incubated with paeonol at a concentration of 47,94,188,376,752 or 1 504 μmol/L for 24 h.② Paeonol down-regulated the transcription level of VEGF in U251 cells in a time-dependent manner.The inhibition rate of VEGF mRNA level was 67.13%,71.33% and 82.04%,respectively,when U251 cells were incubated with paeonol at a concentration of 188 μmol/L for 4,12 or 24 h.③ Paeonol significantly down-regulated the protein expression of VEGF in U251 cells in a dose-dependent manner.The inhibition rate of VEGF protein expression was 33.09%,73.93%,81.23% or 87.85%,respectively,when U251 cells were incubated with paeonol at a concentration of 47,188,376 or 1 504 μmol/L for 24 h.Conclusion Paeonol significantly inhibited the expression of VEGF at mRNA and protein levels in human glioblastoma cells.It may inhibit the angiogenesis of human glioblastoma and increase its chemosensitivity by down-regulating the VEGF expression.